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Phase Separations in Water-Salt Solutions of Polyelectrolyte Complexes Formed by RNA and Polycations: Comparison with DNA Complexes.

Wahlund, Per-Olof LU ; Izumrudov, Vladimir LU ; Gustavsson, Per-Erik LU ; Larsson, Per-Olof LU and Galaev, Igor LU (2003) In Macromolecular Bioscience 3(8). p.404-411
Abstract
Formation of insoluble polyelectrolyte complexes (PECs) between RNA and polycations was followed by measuring the residual RNA absorbance in the solution after separation of the precipitate. The polycations studied were poly(N,N-dimethyldiallylammonium) chloride (pendant type) and 2,5-ionene bromide (integral type) with quaternary amino groups in every monomer unit. The data obtained were compared with the results of analogous studies of DNA-containing PECs. This study is a part of a project aimed at the specific separation of plasmid DNA from RNA, a major problem in the preparative isolation of plasmid DNA. We thus deliberately chose a heterogenous RNA sample as it represents the RNA present in a real cell extract. In contrast to the... (More)
Formation of insoluble polyelectrolyte complexes (PECs) between RNA and polycations was followed by measuring the residual RNA absorbance in the solution after separation of the precipitate. The polycations studied were poly(N,N-dimethyldiallylammonium) chloride (pendant type) and 2,5-ionene bromide (integral type) with quaternary amino groups in every monomer unit. The data obtained were compared with the results of analogous studies of DNA-containing PECs. This study is a part of a project aimed at the specific separation of plasmid DNA from RNA, a major problem in the preparative isolation of plasmid DNA. We thus deliberately chose a heterogenous RNA sample as it represents the RNA present in a real cell extract. In contrast to the exhaustive precipitation of DNA observed at certain values, a significant part of RNA was nonprecipitated at any = [+]/[-], that is, at any ratio of positively charged quaternary amino groups and negatively charged phosphate groups. The addition of sodium chloride increased the nonprecipitated fraction of RNA. DNA, on the other hand, was completely precipitated by both polycations at > 0.7. The less effective precipitation of RNA was probably due to the presence of a considerable fraction of short-chained molecules, incapable of forming a sufficient cooperative system of salt bonds with the polycation. This assumption was supported by a separate experiment, in which the precipitation behavior of RNA fractions of different molecular masses was investigated. The same tendency, while less pronounced, was also ascertained for PECs formed by polycations with DNA fractions of different molecular masses. The possibility of using the revealed differences between DNA and RNA behavior for effective precipitation procedure useful in bioseparation is discussed. The difference in the precipitation behavior of nucleic acids of different molecular masses means there is a possibility for developing an enzymatic assay for DNAase and RNAase activity. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Macromolecular Bioscience
volume
3
issue
8
pages
404 - 411
publisher
John Wiley & Sons
external identifiers
  • wos:000184787300003
  • scopus:0041968489
ISSN
1616-5195
DOI
10.1002/mabi.200350010
language
English
LU publication?
yes
id
03395338-3979-46ab-bedc-f53a2f8feea0 (old id 128638)
date added to LUP
2007-07-03 09:31:42
date last changed
2018-05-29 10:32:52
@article{03395338-3979-46ab-bedc-f53a2f8feea0,
  abstract     = {Formation of insoluble polyelectrolyte complexes (PECs) between RNA and polycations was followed by measuring the residual RNA absorbance in the solution after separation of the precipitate. The polycations studied were poly(N,N-dimethyldiallylammonium) chloride (pendant type) and 2,5-ionene bromide (integral type) with quaternary amino groups in every monomer unit. The data obtained were compared with the results of analogous studies of DNA-containing PECs. This study is a part of a project aimed at the specific separation of plasmid DNA from RNA, a major problem in the preparative isolation of plasmid DNA. We thus deliberately chose a heterogenous RNA sample as it represents the RNA present in a real cell extract. In contrast to the exhaustive precipitation of DNA observed at certain values, a significant part of RNA was nonprecipitated at any = [+]/[-], that is, at any ratio of positively charged quaternary amino groups and negatively charged phosphate groups. The addition of sodium chloride increased the nonprecipitated fraction of RNA. DNA, on the other hand, was completely precipitated by both polycations at > 0.7. The less effective precipitation of RNA was probably due to the presence of a considerable fraction of short-chained molecules, incapable of forming a sufficient cooperative system of salt bonds with the polycation. This assumption was supported by a separate experiment, in which the precipitation behavior of RNA fractions of different molecular masses was investigated. The same tendency, while less pronounced, was also ascertained for PECs formed by polycations with DNA fractions of different molecular masses. The possibility of using the revealed differences between DNA and RNA behavior for effective precipitation procedure useful in bioseparation is discussed. The difference in the precipitation behavior of nucleic acids of different molecular masses means there is a possibility for developing an enzymatic assay for DNAase and RNAase activity.},
  author       = {Wahlund, Per-Olof and Izumrudov, Vladimir and Gustavsson, Per-Erik and Larsson, Per-Olof and Galaev, Igor},
  issn         = {1616-5195},
  language     = {eng},
  number       = {8},
  pages        = {404--411},
  publisher    = {John Wiley & Sons},
  series       = {Macromolecular Bioscience},
  title        = {Phase Separations in Water-Salt Solutions of Polyelectrolyte Complexes Formed by RNA and Polycations: Comparison with DNA Complexes.},
  url          = {http://dx.doi.org/10.1002/mabi.200350010},
  volume       = {3},
  year         = {2003},
}